The ability to engineer a stable cell line that expresses a fluorescent protein under the control of a known promoter sequence provides the unique opportunity to monitor gene promoter activity within living cells through fluorescence intensity measurements. These reporter cells are important tools for assessing cellular response and phenotypic characteristics during in vitro experimental studies. Meaningful quantitative determination of promoter activity from fluorescence intensity measurements requires the development of analytical models that incorporate GFP degradation and productions rates. This project would involve the development of kinetic models and the use of quantitative automated fluorescence microscopy and could result in one or more reporter cell lines that report on cytotoxicity pathways, response to scaffolding for tissue engineered, or cellular differentiation state. Additional applications for engineered cell lines and analytical models include probing the stochastic noise associated with cell signaling pathways and gene expression, and with feedback and feed-forward loops. Combining and correlating multiple readouts within the same cell line is of great interest.
Cell signaling; Fluorescence microscopy; Green fluorescent protein; Image processing; Live cell microscopy; Transfection;